Creating perfused functional vascular channels using 3D bio-printing technology

Abstract

We developed a methodology using 3D bio-printing technology to create a functional in vitro vascular channel with perfused open lumen using only cells and biological matrices. The fabricated vasculature has a tight, confluent endothelium lining, presenting barrier function for both plasma protein and high-molecular weight dextran molecule. The fluidic vascular channel is capable of supporting the viability of tissue up to 5mm in distance at 5 million cells/mL density under the physiological flow condition. In static-cultured vascular channels, active angiogenic sprouting from the vessel surface was observed whereas physiological flow strongly suppressed this process. Gene expression analysis were reported in this study to show the potential of this vessel model in vascular biology research. The methods have great potential in vascularized tissue fabrication using 3D bio-printing technology as the vascular channel is simultaneously created while cells and matrix are printed around the channel in desired 3D patterns. It can also serve as a unique experimental tool for investigating fundamental mechanisms of vascular remodeling with extracellular matrix and maturation process under 3D flow condition.

3D-printed biological organs: medical potential and patenting opportunity

“…3D bioprinting with phase-changing hydrogel [5][6][7][8][9]11,12] --Liquid state, phase-changing hydrogel precursors are printed at high-spatial resolution. --Various types of cells, prepared in a liquid-state suspension, are printed separately and embedded into the hydrogel.…”

“…Therefore, perhaps the most versatile technique could be found from the 'true' 3D bioprinting method, in which the phase-changing hydrogel precursors are printed in a liquid phase as nanoliter-sized liquid droplets, and immediately gelated to maintain the structures in 3D [5]. The method has been used to print > 15 layers of cell-containing hydrogel in an on-demand fashion [6], and has been applied to embed fluidic channels [7] as well as to create vascular networks [8]. The technique is also applied in printing a structure that time-releases soluble cell growth factors to support neural cell differentiation and migration [9].…”

Advances in bioprinted cell-laden hydrogels for skin tissue engineering

“…Promising approaches to generate vascular networks have been reported using inkjet [34,209], laserassisted [203], and extrusion bioprinting [16,92,132]. In general, such approaches are based on four main strategies: (1) direct patterning vascular cells onto a receiving substrate [203], (2) continuous printing of polymeric bioink loaded with endothelial cells followed by polymer removal [96], (3) printing perfusable channels in a 3D construct for subsequent injection of a cell suspension into the empty channel [92], and (4) printing of multicellular spheroids [132]. Vascular cells currently explored in bioprinting include endothelial cells such as human umbilical vein endothelial cells HUVECs or human dermal microvascular endothelial cells (HMVECs), and smooth muscle cells such as human umbilical vein smooth muscle cells.…”

3D Miniaturization of Human Organs for Drug Discovery

“…[88] These vessel generation methods promoted the development of physiologically relevant vascularization and perfusion models. [89,90] Using biological laser printing (BioLP), branch/stem structures of HUVEC and human umbilical vein smooth muscle cells (HUVSMC) were fabricated. [91] The printed structure mimicked vascular networks in tissue and allowed angiogenesis, i.e., the sprouting of new, finer vessels away from the stem Adv.…”

“…[53] In these multicellular aggregates, the need for supporting gels or matrices is eliminated, the adverse effects 3D culture approach for generating a laminated cerebral cortex like structure from pluripotent stem cells. [57,58] Microfabrication Neuroprogenitor cells Microfluidic culture platform containing a relief pattern of soma and axonal compartments connected by microgrooves to direct, isolate, lesion, and biochemically analyze CNS axons [67,68] 3D bioprinting Primary human cortical neurons Discrete layers of primary neutrons in a RGD peptide-modified gellan gum [118][119][120] Intestine (Gut) Self-assembled Stem cells Identified intestinal stem cells and differentiated cells in vitro [59,60] Microfabrication Human epithelial cells Mimic contractility by using mechanochemical actuator [11,19,27,72] Liver Self-assembled Human stem cells 3D culture of self-renewing human liver tissue [61,62] Microfabrication Hepatocytes and fibroblasts Microengineered hepatic microtissues containing hepatocytes and fibroblasts [73][74][75][76][77] 3D bioprinting HepG2 and HUVEC Multilayered organ tissue model [96,[155][156][157] Vessel Microfabrication Rat brain endothelial cells 3D culture in microfluidic device [63][64][65][66] 3D bioprinting HUVECs and HUVSMCs Scaffold-less vessel formation using spheroid fusion [84][85][86][87][88][89][90][91]…”